| Title: |
Effect of protein kinase A activity on the association of ADP-ribosylation factor 1 to Golgi membranes |
| Authors: |
Martín, Maria Esther; Hidalgo, Josefina; Rosa López, José Luis; Crottet, Pascal; Velasco, Ángel |
| Source: |
Articles publicats en revistes (Ciències Fisiològiques) |
| Publisher Information: |
American Society for Biochemistry and Molecular Biology |
| Publication Year: |
2000 |
| Collection: |
Dipòsit Digital de la Universitat de Barcelona |
| Subject Terms: |
Metabolisme; Proteïnes quinases; Aparell de Golgi; Metabolism; Protein kinases; Golgi apparatus |
| Description: |
The small GTP-binding protein ADP-ribosylation factor 1 (ARF1) is an essential component of the molecular machinery that catalyzes the formation of membrane-bound transport intermediates. By using an in vitro assay that reproduces recruitment of cytosolic proteins onto purified, high salt-washed Golgi membranes, we have analyzed the role of cAMP-dependent protein kinase A (PKA) on ARF1 incorporation. Addition to this assay of either pure catalytic subunits of PKA (C-PKA) or cAMP increased ARF1 binding. By contrast, ARF1 association was inhibited following C-PKA inactivation with either PKA inhibitory peptide or RIIalpha as well as after cytosol depletion of C-PKA. C-PKA also stimulated recruitment and activation of a recombinant form of human ARF1 in the absence of additional cytosolic components. The binding step could be dissociated from the activation reaction and found to be independent of guanine nucleotides and saturable. This step was stimulated by C-PKA in an ATP-dependent manner. Dephosphorylated Golgi membranes exhibited a decreased ability to recruit ARF1, and this effect was reverted by addition of C-PKA. Following an increase in the intracellular level of cAMP, ARF proteins redistributed from cytosol to the perinuclear Golgi region of intact cells. Collectively, the results show that PKA exerts a key regulatory role in the recruitment of ARF1 onto Golgi membranes. In contrast, PKA modulators did not affect recruitment of beta-COP onto Golgi membranes containing prebound ARF1. |
| Document Type: |
article in journal/newspaper |
| File Description: |
10 p.; application/pdf |
| Language: |
English |
| Relation: |
Reproducció del document publicat a: https://doi.org/10.1074/jbc.275.25.19050; Journal of Biological Chemistry, 2000, vol. 275, num. 25, p. 19050-19059; https://doi.org/10.1074/jbc.275.25.19050; https://hdl.handle.net/2445/177066; 168288 |
| Availability: |
https://hdl.handle.net/2445/177066 |
| Rights: |
(c) American Society for Biochemistry and Molecular Biology, 2000 ; info:eu-repo/semantics/openAccess |
| Accession Number: |
edsbas.6E9D32DD |
| Database: |
BASE |